A Rapid, Low-Cost Point of Care Diagnostic for detection of Zika virus RNA

用于检测寨卡病毒 RNA 的快速、低成本护理点诊断

• 批准号: 9255921
• 负责人: Sahar Usmani-Brown
• 金额: $ 22.5万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9255921
• 关键词: Acute; Adult; Aedes; African; Amniocentesis; Area; Asians; Bathing; Biological Assay; Blood; Blood specimen; Caribbean region; Central America; Chikungunya virus; Clinical; Colombia; Communities; Complex; Congenital Abnormality; Culicidae; Dengue; Dengue Virus; Dependence; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Disease Outbreaks; Distant; Epidemic; Epidemiologic Monitoring; Equipment; Evaluation; Exanthema; Eye; Female of child bearing age; Fetus; Fever; Flavivirus; Goals; Guillain-Barré Syndrome; Health; Hour; Human; Imagery; Laboratories; Lateral; Mediating; Methods; Microcephaly; Micronesia; Mothers; Newborn Infant; Nucleic Acids; Patients; Performance; Phase; Pregnancy; Pregnant Women; Prenatal care; Procedures; Public Health; Puerto Rico; Quantitative Reverse Transcriptase PCR; RNA; RNA Sequences; RNA-Directed DNA Polymerase; Reaction; Reporting; Research; Resources; Risk; SYBR Green I; Saliva; Sampling; Sensitivity and Specificity; Serologic tests; Serum; Small Business Innovation Research Grant; South America; Southeastern United States; Specificity; Specimen; System; Techniques; Technology; Temperature; Testing; Time; TimeLine; Ultrasonography; United States Dept. of Health and Human Services; Urine; Viral; Viral Genome; Virus; Virus Diseases; Water; West Nile virus; Woman; Work; Zika Virus; assay development; authority; base; chikungunya; clinical Diagnosis; clinical diagnostics; cost; cost effective; cross reactivity; design; diagnostic assay; experience; point of care; point-of-care diagnostics; pregnant; prenatal; prototype; sample collection; screening; transmission process; validation studies; viral RNA; viral detection

ABSTRACT Zika virus (ZIKV) has rapidly emerged and spread through South and Central America, the Caribbean, and Puerto Rico since its last outbreak in Micronesia in 2007. Transmitted by the mosquito Aedes sp, its forecasted spread will have a major impact on the Southeast U.S. Most ZIKV infections remain asymptomatic or present with non-specific rash and fever; therefore, they have been difficult to diagnose and report. However, two major health consequences appear to be associated with the ZIKV outbreak which sets it apart from other flaviviruses such as West Nile Virus and Dengue; namely; (a) transmission from an infected mother to fetus resulting in reports of microcephaly in fetuses; and, (b) Guillain-Barre syndrome (GBS) in adults. Several teams have now developed qRT-PCR assays to detect ZIKV. However such tests are relatively expensive, require well-equipped laboratories with specialized equipment, and the procedure takes at least 3 hours to finish. There is an urgent need for a rapid, sensitive, specific and economical diagnostic test for ZIKV. Such an assay could be routinely used in resource-poor settings as well as in doctors' offices, including as part of regular prenatal care. Therefore, the goal of the SBIR Phase I project is to use loop mediated isothermal nucleic acid amplification (LAMP) to develop a rapid, sensitive point of care diagnostic for ZIKV. This technology is of low complexity, requiring only a water bath. Colorimetric results are visible to the naked eye in one hour or less. We will use serial dilutions of ZIKV and other viruses (including West Nile virus, Dengue viruses, and Chikungunya virus) spiked in human blood, saliva, and urine to determine the assay's sensitivity and specificity. Based on our laboratory's extensive previous experience with LAMP assay development, we forecast a lower limit of detection of 10-100 viral genomes, with very high specificity. In addition to rapid, colorimetric ZIKV detection, we will also develop a rapid, lateral flow assay to facilitate the differential analysis with other flaviviruses. The development of rapid point-of-care assays will reduce the dependence on central laboratory testing facilities for epidemiologic surveillance and clinical diagnosis, a key advantage in the resource-poor areas where the epidemic is currently prevalent. Further, we anticipate that the availability of a rapid test will result in the incorporation of ZIKV testing into the sustainable, routine evaluation of women who are pregnant or anticipating pregnancy, as well as their partners.

抽象的 寨卡病毒（Zikv）迅速出现并通过南美洲，中美洲，加勒比海和 波多黎各自2007年在麦克罗尼亚上次爆发以来。由蚊子SP传播，预测 传播将对美国东南部产生重大影响 非特异性皮疹和发烧；因此，它们很难诊断和报告。但是，两个 重大健康后果似乎与Zikv爆发有关，这使它与其他 黄病毒，例如西尼罗河病毒和登革热；即（a）从感染的母亲到胎儿的传播 导致胎儿小头畸形的报道； （b）成年人的Guillain-Barre综合征（GB）。一些 团队现在已经开发了QRT-PCR分析来检测ZIKV。但是，这样的测试相对昂贵， 需要设备齐全的实验室，并使用专门设备，并且该过程至少需要3个小时才能 结束。迫切需要对ZIKV进行快速，敏感，特定和经济的诊断测试。 这样的测定可以在资源贫乏的设置以及医生办公室中常规使用，包括 定期产前护理。因此，SBIR I期项目的目标是使用循环介导的等温度 核酸扩增（LAMP）为ZIKV开发快速，敏感的护理诊断点。这 技术的复杂性很低，只需要一个水浴。比色的结果可见 一个小时或更短。我们将使用ZIKV和其他病毒的系列稀释液（包括西尼罗河病毒，登革热 病毒和chikungunya病毒）在人，唾液和尿液中尖刺以确定测定的敏感性 和特异性。根据我们实验室以前在LAMP ASSAY开发方面的广泛经验，我们 预测具有非常高的特异性的10-100个病毒基因组的检测下限。除了快速 比色ZIKV检测，我们还将开发快速的侧向流程测定，以促进差分分析 与其他黄病毒。快速护理测定法的发展将减少对中央的依赖 流行病学监测和临床诊断的实验室测试设施，这是 流行病目前普遍存在的资源贫乏地区。此外，我们预计可用性 快速测试将导致将ZIKV测试纳入对妇女的可持续，常规评估中 怀孕或预期怀孕及其伴侣。